caa a22 4500 463238338 CHVBK 20180405153305.0 cr unu---uuuuu 170326e20070501xx s 000 0 eng 10.1007/s11259-007-3470-1 doi (NATIONALLICENCE)springer-10.1007/s11259-007-3470-1 Development of a Simple Enzyme Immunoassay for the Determination of Ovine Luteinizing Hormone [Elektronische Daten] [J. Valares, J. Abecia, F. Forcada, I. Palacín, L. Mata, P. Razquin] The present study describes the development and validation of a simple sensitive and specific sandwich enzyme immunoassay (EIA) for the quantification of ovine luteinizing hormone (LH) in plasma. Microtitre plates were coated with the capture antibody 518b7 anti-bovine LH. A second peroxidase-labelled anti-ovine LH antibody was used as tracer. A simple 3-step procedure was used for the sample analysis; (1) incubation of standards and samples with the pre-coated antibody plates for 2 h at 37°C; (2) incubation with the peroxidase-labelled antibody for 1 h at room temperature; and (3) colour development with TMB substrate. A linear dose-response curve was obtained in the range 0-10 ng/ml (r 2 > 0.99). The detection limit was 0.05 ng/ml, and the intra-assay and inter-assay coefficients of variation were 7% and 11.7%, respectively. The theoretical stability of microplates and reagents was calculated, this being greater than one year. Low or undetectable cross-reactivities were recorded for follicle-stimulating hormone, bovine thyroid-stimulating hormone, equine chorionic gonadotrophin and a gonadotrophin-releasing hormone (GnRH) analogue. The EIA was biologically validated by the determination of plasma LH concentrations of nine Rasa Aragonesa ovariectomized and estradiol-implanted ewes after a double GnRH challenge. In conclusion, this enzyme immunoassay provides an efficient, simple and sensitive method for the routine analysis of ovine LH. Springer Science + Business Media, Inc., 2007 ELISA nationallicence luteinizing hormone nationallicence sheep nationallicence E2 : estradiol nationallicence eCG : equine chorionic gonadotrophin nationallicence EIA : enzyme immunoassay nationallicence ELISA : enzyme-linked immunosorbent assay nationallicence FSH : follicle-stimulating hormone nationallicence GnRH : gonadotrophin-releasing hormone nationallicence HRP : horseradish peroxidase nationallicence LH : luteinizing hormone nationallicence OVX : ovariectomized nationallicence RIA : radioimmunoassay nationallicence TMB : tetramethylbenzidine liquid substrate system nationallicence TSH : thyroid-stimulating hormone nationallicence Valares J. Departamento de Producción Animal y Ciencia de los Alimentos, Facultad de Veterinaria de Zaragoza, Zaragoza aut Abecia J. Departamento de Producción Animal y Ciencia de los Alimentos, Facultad de Veterinaria de Zaragoza, Zaragoza aut Forcada F. Departamento de Producción Animal y Ciencia de los Alimentos, Facultad de Veterinaria de Zaragoza, Zaragoza aut Palacín I. Departamento de Producción Animal y Ciencia de los Alimentos, Facultad de Veterinaria de Zaragoza, Zaragoza aut Mata L. Zeu Inmunotec, Zaragoza, Spain aut Razquin P. Zeu Inmunotec, Zaragoza, Spain aut Veterinary Research Communications Kluwer Academic Publishers 31/4(2007-05-01), 427-436 0165-7380 31:4<427 2007 31 11259 https://doi.org/10.1007/s11259-007-3470-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11259-007-3470-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Valares J. Departamento de Producción Animal y Ciencia de los Alimentos, Facultad de Veterinaria de Zaragoza, Zaragoza aut NATIONALLICENCE 700 1- Abecia J. Departamento de Producción Animal y Ciencia de los Alimentos, Facultad de Veterinaria de Zaragoza, Zaragoza aut NATIONALLICENCE 700 1- Forcada F. Departamento de Producción Animal y Ciencia de los Alimentos, Facultad de Veterinaria de Zaragoza, Zaragoza aut NATIONALLICENCE 700 1- Palacín I. Departamento de Producción Animal y Ciencia de los Alimentos, Facultad de Veterinaria de Zaragoza, Zaragoza aut NATIONALLICENCE 700 1- Mata L. Zeu Inmunotec, Zaragoza, Spain aut NATIONALLICENCE 700 1- Razquin P. Zeu Inmunotec, Zaragoza, Spain aut NATIONALLICENCE 773 0- Veterinary Research Communications Kluwer Academic Publishers 31/4(2007-05-01), 427-436 0165-7380 31:4<427 2007 31 11259 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer